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Journal of Taibah University for Science 10 (2016) 497–504

Production of biogas from poultry litter mixed with the co-substrate

cow dung
Mohammad Roman Miah a , Abul Kalam Md. Lutfor Rahman a,∗ ,
Muhammad Rajibul Akanda a , Abdullah Pulak a , Md. Abdur Rouf b
a Department of Chemistry, Jagannath University, Dhaka 1100, Bangladesh
b Institute of Fuel Research and Development, BCSIR, Dr. Qudrat-I-Khuda Road, Dhanmondi, 1205 Dhaka, Bangladesh
Received 23 March 2015; received in revised form 4 July 2015; accepted 31 July 2015
Available online 6 November 2015

Abstract
Poultry litter (a mixture of rice hulls, sawdust and chicken excreta of broilers) mixed with the co-substrate cow dung and poultry
droppings was evaluated under anaerobic conditions for the production of biogas (methane). Four laboratory scale reactors, R1,
R2, R3 and R4, were set up with different proportions of waste poultry litter, cow dung and poultry droppings and had a 6% total
solid concentration. Digestion was carried out for 50 days at room temperature, 32 ± 3 ◦ C. Volatile solid degradation and specific
gas production in the four reactors was 46%, 51.99%, 51.96%, 43% and 0.263, 0.469, 0.419, 0.221 l/g, respectively, based on the
volatile solid (VS) feed. The methane yields were 71%, 72.5%, 72.6% and 70%, respectively. The COD reductions were 46.1%,
50.76%, 48.23% and 45.12%, respectively. A kinetic analysis showed that the anaerobic digestion of poultry litter with a co-substrate
followed first order kinetics. Among the experimental reactors, R2 (25% cow dung, 75% poultry litter) gave the optimum results: a
VS reduction of 51.99%, a specific gas yield of 0.469 l/g and a methane yield of 72.5%.
© 2015 The Authors. Production and hosting by Elsevier B.V. on behalf of Taibah University. This is an open access article under
the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Keywords: Poultry litter; Co-substrate; C/N ratio; Anaerobic digestion; Biogas

1. Introduction produced by the biomethanation process, and the effluent

Biogas is an environmental friendly and one of the
most efficient and effective options for renewable energy
among various other alternative sources [1]. Biogas is
∗ Corresponding author. Tel.: +880 1732108451.
E-mail address: lrahman1973@gmail.com (A.K.Md.L. Rahman).
Peer review under responsibility of Taibah University.
from the process is rich in essential nutrients that can
be utilized as a very good fertilizer. Biomethanation is
the degradation of organic materials by microorganisms
in the absence of oxygen. It is a multi-step biological
process in which organic carbon is converted mostly to
carbon dioxide and methane [2]. Biogas can be produced
from variety of substrates, such as animal manure, energy
crops, industrial waste and so on. The typical reactions
that occur in the anaerobic digestion process [1,3] are:
Acetogenic bacteria:
Cellulose C6 H12 O6 → 2CH3 COOH + 2CO2 + 4H2

http://dx.doi.org/10.1016/j.jtusci.2015.07.007
1658-3655 © 2015 The Authors. Production and hosting by Elsevier B.V. on behalf of Taibah University. This is an open access article under the
CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
498 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504
Methanogenic bacteria:
CH3 COOH → CH4 + CO2 , CO2 + H2 → CH4
Bangladesh is home to approximately 160 million
people. Recently, poultry farms in Bangladesh have been
rapidly expanding to meet the growing demand for meat
and eggs. There are 123 million chickens [4] and approx-
imately 50,000.00 poultry farms [5] in Bangladesh. It
is estimated that approximately 1,560,000.00 metric
tonnes of poultry manure is produced in Bangladesh
each year. Including bedding material, wasted feed and
feathers, the total poultry waste (litter) is much higher.
Most of the waste materials are dumped into nearby sites,
although a small portion of poultry waste is used for fish
and crop production by farmers. Crude dumping of this
waste is not only unattractive but also environmentally
unsafe. Biogas production can be a sustainable solution
to treat waste materials, and the cost of waste treatment
using this process is low. Poultry waste, cow dung and
other waste have been used for biogas production [1,6],
but the efficiency of the gas production is low.
In this study, biomethanation of poultry litter was
studied with the co-substrates cow dung and poultry
droppings. A comparative analysis using different mate-
rials was performed on biogas production.
2. Materials and methods
2.1. Sample collection
The poultry litter and poultry droppings were col-
lected from Joypara, Dohar, Dhaka. Cow dung was also
used in various proportions as a co-substrate to maintain
the C/N ratio.
2.2. Preparation of reactors
The anaerobic digestion process was studied in batch
reactors to develop an appropriate technology for the
production of biogas from the solid waste from poultry
Fig. 1. Outline of the experimental setup for biogas generation.
farms. A known amount of substrate containing a mix-
ture of waste was transferred into a 2-litre, wide mouth
glass bottle. All of the bottles were sealed with air tight
rubber stoppers, and another bottle was filled with water
to collect gas and was equipped with glass tubes for
gas removal. Biogas produced by anaerobic digestion
was collected by the water displacement method [7].
Four reactors were prepared using various proportions
of poultry litter and other materials. Reactor R1 was set
up using poultry litter alone. R2 was prepared using 75%
poultry litter and 25% cow dung, R3 was 50% poultry
litter and 50% cow dung and R4 was 70% poultry lit-
ter and 30% poultry droppings. A reactor is pictured in
Fig. 1.
2.3. Sample analysis
The sample pH was measured with a digital pH metre
(HANNA, HI 98204). The carbon and nitrogen con-
tent of the poultry litter, poultry droppings and cow
dung were determined with a C–H–N elemental anal-
yser. Total solids (TS) were determined by incubating a
sample at 104 ◦ C until no further weight change was evi-
dent, and volatile solids (VS) were measured by the loss
on ignition of the dried sample at 550 ◦ C. The composi-
tion of the gas was measured using an Orsat gas analyser.
COD was determined by a chemical method, and the
calorific value was determined in a bomb calorimeter.
The total gas production was measured via the water
displacement method at an interval of 24 h. The pH, VS
reduction and COD reduction were measured every 10
days throughout the experiment. Each experiment was
conducted at a temperature of 32 ± 3◦ C for 50 days.
3. Results and discussion
Various parameters of the raw materials are shown in
Table 1. Table 1 shows that the C/N ratio of poultry litter
is 7.5, which is quite low for optimum biogas generation
[8], and it can be increased by mixing with cow dung
 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504 499

Table 1
Characterization of raw materials used in the biogas digester.
Constituents Poultry litter Poultry dropping Cow dung Inoculums

pH (6% of slurry) 8.5 7.0 6.3 6.43

Moisture content (%) 27.68 77.5 83 96
TS (%) 72.32 22.5 17 6
VS (%) 83.95 66.72 89 88.66
COD (g/l) 76.5 65.3 – –
C/N ratio 7.5 6.5 24 –
Calorific value (kcal/kg) 4519.77 3980.75 4658.07 –

DS, desulfurized slag; WQS, water-quenched slag; AAR, arsenic–alkali residue; BFD, blast furnace dust.

because cow dung has a high C/N ratio. When a substrate
with a low C/N ratio is mixed with a substrate that has a
high C/N ratio, better performance is achieved [7]. The
pH of the poultry litter is 8.5. The buffering capacity of
cow dung is very good, and it also acts as the source of
various microorganisms required for anaerobic digestion
[9].
3.1. Biomethanation potential of digesters
Cumulative gas generation in different reactors is
plotted in Fig. 2. The gas generation rate was higher
in reactor R2 than in the others, even though reactor R2
contained less cow dung than R3. It can be concluded
that the potential gas generation from poultry litter via
anaerobic digestion with a certain proportion of added
cow dung is high.
3.2. Effect of volatile solids on gas production
Gas accumulation was observed daily in reactors con-
taining different amounts of poultry litter. The daily gas
accumulation in the different reactors was used to deter-
mine the peak gas production as related to the initial
volatile solid concentration. The relationship between
periods of peak gas production and the initial volatile
solid concentration in the different reactors is shown in
Fig. 3, which shows that peak gas generation occurred
earlier when the volatile solids concentration was higher.
The lower the initial VS concentration, the more time
required for peak gas production in the reactor.
Cumulative degradation of VS and cumulative gas
production with time are shown in Fig. 4 denoted by
(a), (b), (c) and (d) for reactors R1, R2, R3 and R4,
respectively. Gas generation from the degradation of the
VS was 0.57, 0.902, 0.808 and 0.52 l/g, respectively. The
biogas yields were comparable with the trends reported
by Thangamani and co-workers [9]. The gas generation
from the degraded volatile solids was higher in R2 than
in the other three reactors.
The relationship between the percent of volatile solids
(VS) reduction and the gas yield is shown in Table 4. The
values for VS destruction indicate that the gas yield in
R1 (which contained 100% poultry litter) was lower than
R2 and R3 and was higher than R4. Because there was
no cow dung in R1 and R4 to stimulate the process with
bacteria, poultry litter alone was the major source of bio-
gas in the slurry. Therefore, VS reduction was lower in

30000 40
Cumulative gas production, mL

Observed peak gas generation

34
R2 30
R3 30 27
20000
24
(days)

20 R² = 0.993
R1
10000 R4
10

0
0 48 49 50 51 52 53
0 20 40 60 Initial volatile solid conc. (g/l)
Time, Days
Fig. 3. Observed peak gas generation time with initial volatile solid
Fig. 2. Cumulative gas production with time at different reactors. conc. (g/l).
500 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504

30 30 30 30

Degraded Volatile solids (g/l)

Degraded Volatile Solids(g/l)

Degraded Volatile solids (g/l)

Degraded Volatile Solids(g/l)

Cumulative gas (l)

25 Cummulative Gas(l) 25 25 Cumm ulative Gas(l) 25

R2

Cumulative gas (l)

20 20 20 20

15 15 15 15

10 10 10 10

5 R1 5 5 5

0 0 0 0
0 20 40 60 0 20 40 60
Time,days Time, Days

30 30

Degraded Volatile solids (g/l)

30 30
Degraded Volatile solids (g/l)

Degraded volatile solids(g/l)

25 25 Degraded volatile solids(g/l)

Cumulative gas (liter)

Cummulative gas(l) 25 25

Cumulative gas (l)

Cummulative gas(l)
20 20 20 20

15 15 15 15

10 10 10 10
R3
5 5 5 5
R4
0 0
0 0
0 20 40 60
0 20 40 60
Time,days Time,days

Fig. 4. Cumulative destruction of volatile solid (VS) and cumulative gas production with time for reactors R1, R2, R3 and R4.

both reactors. On the other hand, in R2, which contained 90 30

25% cow dung and 75% poultry litter, the volatile solids COD Value, g/l

Cumulative gas production(l)

81 27
reduction was 51.99%. Although gas production in R3 Cumulative gas production (litre)
COD value decrease(g/l)

72 24
was lower than in R2, it was comparable, which indi-
63 21
cates that VS reduction not only depends on the initial
concentration but also on the type of volatile matter in 54 18
the slurry. The yield of gas generation was higher in R2 45 15
than in the other three reactors. These results imply that 36 12
poultry litter alone is not a suitable option for optimum 27 9
gas generation.
18 6
9 3
3.3. Effect of COD reduction on cumulative gas
0 0
production
0 6 12 18 24 30 36 42 48 54 60
Time, days
The chemical oxygen demand (COD) of the slurry
was considerably reduced by the anaerobic digestion Fig. 5. COD reduction and cumulative gas production with time in R2
process. The reduction of the COD implies the reduction reactor.
of the pollutant load from any substrate during the treat-
ment process. The COD reduction and cumulative gas 3.4. Effect of cow dung
production for R2 is shown in Fig. 5. For reactors R1, R2,
R3 and R4, the COD reduction was 46.1, 50.76, 48.23 The volume of gas production with different percent-
and 45.12%, respectively. The maximum COD reduction ages of cow dung in the slurry in different reactors is
was achieved in the R2 reactor in which the maximum shown in Fig. 6. Because the initial pH of the poultry
amount of gas was produced. The COD reduction was litter was high, cow dung also acted as a buffer and a
comparable with the reference value given by Rahman seeding material. No cow dung was present in reactor
and Muyeed [3]. R1; therefore, the onset of minimum gas generation in
 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504
30
25 R2
R3
Gas Production(l)
23.99
20 21.78
13.24
15
R1
10 Gas production(litre)
5 in R1, a considerable amount of gas was obtained. This
0
0% 20% 40%
% of cowdun g used
Fig. 6. Effect of cow dung on gas production.
this reactor was at 4 days because more time was required
to digest the poultry litter in the absence of cow dung.
The data from the four reactors implies that optimum gas
generation was obtained when 25% of the total solids
in the slurry was cow dung. The percentages of C and
N were 31.6 and 4.21, respectively. The value of the
C/N ratio was 7.5 for broiler poultry litter and 24 for
cow dung. Cow dung contains much less nitrogen than
poultry litter. Therefore, the C/N ratio increases when
poultry litter is mixed with cow dung. The value of C/N
ratio should be more than 10 or ammonia toxicity may
develop because of the high concentration of N. A C/N
near 20 is considered to be optimum [7]. It is assumed
that an increase in cow dung increases the C/N ratio and
consequently decreases the protein content. Anaerobic
digestion might not be favoured under such conditions.
For this reason our experiment indicated that 25% cow
dung and 75% poultry litter is the optimum composition.
The biogas yield at various C/N ratios in the four
reactors is shown in Fig. 7. The C/N ratio of the poultry
litter was very low, only 7.5. Generally, a low C/N ratio
indicates that a substrate has a high protein content. In
Fig. 7. Effect of C/N ratio against the yield of biogas.
501
an anaerobic process, the excess protein is degraded to
NH3 , and an elevated concentration of NH3 may be toxic
to anaerobic bacteria. For this reason, cow dung was used
as a co-substrate to maintain the C/N ratio. The reactors
were operated at C/N ratios of 7.5, 17, 20.88 and 6.99,
which were slightly lower than the reference value of
20–30 [7]. The highest amount of biogas was obtained
at a C/N ratio of 17 in the R2 reactor. The reactors oper-
ated without any setback; even at the lowest C/N value
could be because poultry litter is nutrient-rich and con-
tains adequate amounts of carbon, oxygen, hydrogen,
60%
nitrogen, phosphorous, potassium, calcium, and mag-
nesium as well as a number of trace elements that are
essential for the growth of anaerobic bacteria.
3.5. Monitoring of pH
The pH is another important parameter that must be
monitored in an anaerobic digestion process. According
to Rahman and Muyeed [3], the desirable pH range for
an anaerobic digestion process is 6.5–8.0, and the high-
est gas yield was observed by the Chengdu Research
Institute at a pH of 7.5–8.0. The pH ranges in R1, R2,
R3 and R4 were 8.5–7.3, 7.6–7.2, 6.9–7.15 and 8.6–7.8,
respectively.
3.6. Biodegradability of the feed mixture
The refractory fraction in the feed mixture is the
portion of the initial VS that remains in the digester as
the solid retention time (SRT) approaches infinity [8,10]
and is an indicator of the extent of the biodegradability of
the substrate. It is essential to monitor the biodegradable
fraction of volatile solids in the feed to maintain better
operational control of the process. The refractory frac-
tion can be determined graphically from the intercept of a
plot of (S/S0 ) and (S0 *t)−1 [9], where S = substrate con-
centration (g/l), S0 = initial VS concentration (g/l) and
t = retention time (d). The biodegradability of the feed
mixture was determined from the intercept and was in the
range of 47–56% of the influent volatile solids concentra-
tion shown in Table 2. The graphs used to determine the
refractory fraction in the reactors are shown in Fig. 8. The
biodegradability factor obtained from the graph indicates
that the presence of resistant volatile matter is the major
portion of volatile solids in the digester. This reasonably
conforms to the experimentally determined VS destruc-
tion efficiency of 43–52%. The biodegradable fraction
of volatile solids obtained from the graphical model and
the experimental values are presented in Table 2.
502 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504

Table 2
Refractory fraction, biodegradable fraction and total VS destruction observed in different reactors.
Reactor Refractory % of VS destruction based on % of VS destruction
fraction of VS (%) refractory fraction of VS based on total of VS

R1 52.6 47.4 46
R2 45.2 54.8 51.99
R3 44.2 55.8 51.96
R4 55.7 44.3 43

4. Kinetic analysis (l), V is the volume of the reactor (l) and C is the yield
constant (l/g).
An anaerobic digestion process is generally described For a batch reactor, the substrate remaining in the
by a first order kinetic model, which is based on two fac- digester is given by integrating Eq. (1),
tors: the rate of substrate conversion to biogas is directly
proportional to the substrate concentration, and the vol- S = S0 exp(−k(t − t0 )); t  t0 . (3)
ume of gas generated is proportional to the mass of the where t0 is the lag time (d). This model [12,13] describes
substrate degraded [11]. the behaviour of an average reactor with a longer reten-
The corresponding kinetic equations are: tion time. From Eqs. (2) and (3), the cumulative gas
dS production can be predicted by,
= −kS, (1)
dt G = CVS0 [1 − exp(−k(t − t0 ))]. (4)
G = CV (S − S0 ). (2)
Rearranging by taking natural logarithm gives,
where S is the final substrate concentration (g/l), S0 is the  
1−G
initial substrate concentration (g/l), k is the rate constant, ln = −kt + kt0 . (5)
t is the time (days), G is the cumulative gas production CVS0

1.2
1.2
1 R1
1 R2
0.8 0.8
S/So

S/So

0.6 0.6
y = 223.9x + 0.526
R² = 0.840
0.4 0.4
y = 236.4x + 0.452
0.2 0.2 R² = 0.876
0 0
0 0.001 0.002 0.003 0 0.001 0.002 0.003
1/(So*t)
1/(So*t)

1.2 1.2
R4
1 R3 1

0.8 0.8
S/So
S/So

0.6 0.6
0.4
0.4 y = 212.8x + 0.557
y = 248.7x + 0.442
R² = 0.898 0.2 R² = 0.828
0.2
0
0
0 0.001 0.002 0.003
0 0.001 0.002 0.003
1/(So*t) 1/(So*t)

Fig. 8. Refractory fraction in four different reactors R1, R2, R3 and R4.
 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504 503

Time, days Time, days

0 20 40 60 0 20 40 60
0 0
-0.1 -0.1
R1
R2
-0.2 -0.2

ln(1-G/CVSo )
ln(1-G/CVSo )

-0.3 -0.3
-0.4 -0.4
-0.5 -0.5
y = -0.013x + 0.078
-0.6 R² = 0.998 -0.6
y = -0.015x + 0.023
-0.7 -0.7 R² = 0.999
-0.8 -0.8

Time, days Time, days

0 20 40 60 0 20 40 60
0 0
-0.1 -0.1
R4
-0.2 R3 -0.2
ln(1-G/CVSo )

ln(1-G/CVSo )
-0.3 -0.3
-0.4 -0.4
-0.5 -0.5 y = -0.012x + 0.076
-0.6 R² = 0.995
y = -0.015x + 0.021 -0.6
-0.7 R² = 0.997 -0.7
-0.8 -0.8

Fig. 9. Logarithmic plot of gas production with time for different reactors experimented.

A graphical analysis of the kinetic model for the data
from R1, R2, R3 and R4 are shown in Fig. 9 in which
ln(1 − G/CVS0 ) versus time has been plotted for the four
reactors. The rate constant and lag time were determined
by using Eq. (5).
The rate constant and lag time can be calculated from
the kinetic analysis for four different reactors from the
yield constant (C), the slope of −kt and the intercept of
kt0 . The yield constant, rate constant and lag time for
various initial VS concentrations are shown in Table 3,

Table 3
Yield constant, rate constant, correlation coefficient, predicted and observed lag time for various reactors.
Reactor Yield constant, Rate constant, Correlation coefficient, Lag time, t0 days Lag time, t0 days
l/g k R2 (predicted) (observed)

R1 0.57 0.013 0.998 7.8 4

R2 0.902 0.015 0.999 2.3 2
R3 0.808 0.015 0.997 2.1 2
R4 0.52 0.012 0.995 7.6 5

R1 = 100% poultry litter; R2 = 75% poultry litter and 25% cow dung; R3 = 50% poultry litter and 50% cow dung; and R4 = 70% poultry litter and
30% poultry dropping.

Table 4
Yield of gas generation with % of VS destruction among four reactors.
Reactors VS (g/l) VS reduction (%) Yield of gas
generation with VS
Initial Final destruction (l/g)
R1 50.37 27.14 46 0.57
R2 51.15 24.56 51.99 0.902
R3 51.9 24.94 51.96 0.808
R4 47.24 26.93 43 0.52
504 M.R. Miah et al. / Journal of Taibah University for Science 10 (2016) 497–504
which indicates that the observed lag time is more or
less in the same range as predicted by using Eq. (5). For
all of the reactors, a logarithmic plot of gas production
and time was linear with a negative slope. The results
fitted a first order kinetic model well. The maximum
yield constant was found in reactor R2 in which 75%
poultry litter with 25% cow dung was used.
5. Conclusions
Poultry litter from broilers mixed with an optimum
proportion of cow dung was found to be a substrate
with a high potential for biogas generation by anaero-
bic digestion. Batch reactors R1, R2, R3 and R4 were
operated with an initial volatile solid concentration of
50.37, 51.15, 51.9 and 47.24 g/l, respectively, and the
corresponding specific gas production obtained in terms
of the volatile solids feed were 0.263, 0.469, 0.419
and 0.221 l/g, respectively. The VS degradation efficien-
cies were 46, 51.99, 51.96 and 43%, respectively. The
methane content in the biogas generated from different
reactors varied from 70 to 72.6%. Kinetic analyses of the
data for the reactors indicated that a first order kinetic
model was adequate to describe the anaerobic digestion
of broiler poultry litter in the presence of co-substrates.
Based on the volatile solids degradation efficiencies and
specific gas production in terms of the quantity of volatile
solid feed, it was confirmed that poultry litter (broiler)
is desirable for anaerobic digestion when mixed with
a specific proportion of cow dung to obtain a bench-
mark quantity of biogas (methane). Cow dung at 25%
and poultry litter at 75% (in the R2 reactor) was the
optimum composition for biogas production.
Acknowledgments
This work was supported by the Ministry of
Science and Technology, Bangladesh, under the
project “Direct catalytic conversion of biogas to
methanol” (No. 39.009.006.01.00.049/2013-2014/EAS-
15/292; Date: 06/11/2013). We would like to acknowl-
edge the help from Department of Chemistry, Jagannath
University and BCSIR.
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